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Updated: Sep 26, 2026

In Vivo Imaging of Cerebrospinal Fluid Transport through the Intact Mouse Skull using Fluorescence Macroscopy
Published on: July 29, 2019
Simultaneous real-time imaging of upper cervical cerebrospinal fluid and vascular dynamics using ultrafast
Amir H Shaker1,2,3, Amir H G Arani1,2, Bryn A Martin4,5
1Mallinckrodt Institute of Radiology, Washington University School of Medicine, St. Louis, MO, United States.
Abstract:
Cerebrospinal fluid (CSF) circulation is tightly coupled to cerebral blood flow under the fixed-volume constraint of the cranial vault, with cardiac pulsations and respiration acting as dominant physiological drivers. Disruption of these flow dynamics has been implicated in various neurological disorders, motivating the need for imaging methods that capture CSF and vascular flow simultaneously and in real time. Conventional phase contrast MRI (PC-MRI) provides quantitative CSF velocity measurements but relies on cardiac gating and velocity encoding, which limit temporal resolution, dynamic range, and sensitivity to non-cardiac fluctuations. Here, we implement a single-slice ultrafast inflow-weighted echo-planar imaging (EPI), in which signal intensity reflects the replacement of saturated spins by freshly inflowing, unsaturated water molecules, yielding higher signal with higher flow velocity. This allows simultaneous assessment of arterial inflow, venous outflow, and CSF motion at high temporal resolution. Ultrafast EPI was acquired at the C2-C3 spinal level, enabling real-time imaging of CSF flow dynamics in the cervical spinal canal alongside arterial and venous blood flow in major cervical vessels (frame rate: 21.7 Hz). Furthermore, intrinsic arterial signal fluctuations were leveraged as a timing reference to reconstruct cardiac-resolved CSF dynamics for individual cardiac cycles without external physiological recordings. Frequency-domain analysis revealed distinct spectral signatures in CSF flow compared with neurovascular flow, including broadened cardiac peaks and enhanced respiratory modulation, particularly within the ventral spinal canal. In contrast, dorsal CSF showed increased power within the cardiac frequency band, higher coherence with cervical vasculature at the cardiac frequency, and reduced non-cardiac contributions. Time-domain analysis showed strong correlation between CSF flow waveforms derived from ultrafast EPI and PC-MRI. Beyond ensemble-averaged waveforms, ultrafast EPI enabled beat-to-beat analysis, revealing substantial cycle-to-cycle variability in CSF flow that is not captured by time-averaged gated approaches. Together, these findings establish ultrafast EPI as a rapid, complementary framework to PC-MRI, enabling real-time neurofluid imaging with integrated time- and frequency-domain characterization of CSF and neurovascular flow dynamics.
